RCC2 Expression Stimulates ER-Positive Breast Tumorigenesis
RCC2 Expression Stimulates ER-Positive Breast Tumorigenesis
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RCC2 表达刺激 ER 阳性乳腺肿瘤发生
DOI：
10.1155/2020/5619462
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发表时间：
2020-05-23
影响因子：
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通讯作者：
Chang, Xiaotian
中科院分区：
文献类型：
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作者：
Wang, Weiqi;Xu, Bing;Chang, Xiaotian
目的染色体凝集调节因子2（RCC 2）参与细胞分裂的调控。本研究探讨了RCC 2表达对乳腺肿瘤发生的影响。方法用抗RCC 2 siRNA或RCC 2表达质粒转染雌激素受体阳性（ER+）乳腺癌MCF-7细胞。分别用CCK-8法、伤口愈合法和流式细胞术检测转染细胞的增殖、凋亡、迁移和细胞因子的产生。用PCR芯片检测转染抗RCC 2 siRNA的MCF-7细胞中RCC 2的致瘤通路。MCF-7细胞也用含有抗RCC 2短发夹RNA的慢病毒转染，并注射到BALB/c裸鼠中以产生荷瘤小鼠。使用磁共振成像通过扩散加权成像分析检查小鼠模型中的肿瘤生长。结果与乳腺纤维腺瘤组织相比，ER阳性乳腺癌组织中RCC 2的表达明显增加。在MCF-7细胞中，抑制RCC 2表达可降低细胞迁移并刺激凋亡，而过表达RCC 2可刺激细胞迁移并抑制凋亡。抑制RCC 2表达显著降低了荷瘤小鼠的乳腺肿瘤生长和IL-6水平。PCR芯片显示，抑制RCC 2表达可显著降低MCF-7细胞中两个已知的肿瘤增强基因IGF 1和TWIST 1的表达;相反，过表达RCC 2可增加这两个基因在转染细胞中的表达水平。在MCF-7细胞中抑制RCC 2表达后，在小鼠模型中验证了该结果。此外，雌二醇-17 β抑制MCF-7细胞凋亡，刺激细胞增殖和细胞迁移，并增加RCC 2、IGF 1和TWIST 1表达。siRNA介导的RCC 2表达抑制减轻了雌激素对MCF-7细胞凋亡的抑制作用，而过表达RCC 2增强了雌激素驱动的凋亡抑制作用。在存在或不存在雌二醇-17 β的情况下，修饰RCC 2表达对MCF-7细胞增殖没有影响。结论雌激素诱导的RCC 2表达促进IGF 1、TWIST 1和IL-6表达，刺激细胞迁移，抑制细胞凋亡，从而促进ER +乳腺肿瘤的发生。
Objective Regulator of chromosome condensation 2 (RCC2) has been reported to be involved in the regulation of cell cleavage. This study investigated the effect of RCC2 expression on breast tumorigenesis. Methods MCF-7 cells originating from estrogen receptor-positive (ER+) breast cancer were transfected with anti-RCC2 siRNA or RCC2-expressing plasmids. Cell proliferation, apoptosis, migration, and cytokine production in the transfected cells were examined using the CCK-8 assay, wound healing assay, and flow cytometry, respectively. PCR array was used to investigate the tumorigenic pathway of RCC2 in MCF-7 cells transfected with the anti-RCC2 siRNA. MCF-7 cells were also transfected with lentivirus-containing anti-RCC2 short hairpin RNA and were injected into BALB/c nude mice to generate tumor-bearing mice. Tumor growth in the mouse model was examined using magnetic resonance imaging by diffusion-weighted imaging analysis. Results Western blotting and immunohistochemistry detected significantly increased expression of RCC2 in ER + breast tumor tissues compared with breast fibroadenoma samples. Inhibiting RCC2 expression decreased cell migration and stimulated apoptosis in MCF-7 cells, while overexpressing RCC2 stimulated cell migration and inhibited apoptosis. The inhibition of RCC2 expression significantly decreased breast tumor growth and IL-6 levels in the tumor-bearing mice. PCR array demonstrated that inhibiting RCC2 expression significantly decreased the expression of IGF1 and TWIST1, two well-known tumor-enhancing genes, in MCF-7 cells; conversely, overexpressing RCC2 increased the expression levels of these two genes in the transfected cells. This result was verified in the mouse model following inhibition of RCC2 expression in MCF-7 cells. Additionally, estradiol-17β suppressed MCF-7 cell apoptosis, stimulated cell proliferation and cell migration, and increased RCC2, IGF1, and TWIST1 expression. The siRNA-mediated inhibition of RCC2 expression alleviated the inhibitory effects of estrogen on apoptosis in MCF-7 cells, while overexpressing RCC2 enhanced the estrogen-driven inhibition of apoptosis. Modifying RCC2 expression had no impact on MCF-7 cell proliferation in the presence or absence of estradiol-17β. Conclusions Our results suggest that estrogen-induced RCC2 expression prompts IGF1, TWIST1, and IL-6 expression, stimulates cell migration, and inhibits apoptosis to contribute to ER + breast tumorigenesis.